ABSTRACT
GalNAc oligonucleotide conjugates demonstrate improved potency in vivo due to selective and efficient delivery to hepatocytes in the liver via receptor-mediated endocytosis. GalNAc-siRNA and GalNAc-antisense oligonucleotides are at various stages of clinical trials, while the first two drugs were already approved by FDA. Also, GalNAc conjugates are excellent tools for functional genomics and target validation in vivo. The number of GalNAc residues in a conjugate is crucial for delivery as cooperative interaction of several GalNAc residues with asialoglycoprotein receptor enhances delivery in vitro and in vivo. Here we provide a robust protocol for the synthesis of triple GalNAc CPG solid support and GalNAc phosphoramidite, synthesis and purification of RNA conjugates with multiple GalNAc residues either to 5'-end or 3'-end and siRNA duplex formation.
Subject(s)
Acetylgalactosamine/chemical synthesis , Immobilized Nucleic Acids/chemical synthesis , Oligodeoxyribonucleotides/chemical synthesis , Organophosphorus Compounds/chemical synthesis , RNA, Small Interfering/chemical synthesis , Acetylgalactosamine/analogs & derivatives , Research Design , WorkflowABSTRACT
Patients whose cancer is detected early are much more likely to have a positive prognosis and outcome. Nanoflares hold promise as a practical diagnostic platform for the early detection of cancer markers in living cells. These probes are based on spherical nucleic acid (SNAs) and are typically composed of gold nanoparticle cores and densely packed and highly oriented oligonucleotide shells; these sequences are complementary to specific mRNA targets and are hybridized to fluorophore-labeled reporter strands. Nanoflares take advantage of the highly efficient fluorescence quenching properties of gold, the rapid cellular uptake of SNAs that occurs without the use of transfection agents, and the enzymatic stability of such constructs to report a highly sensitive and specific signal in the presence of intracellular target mRNA. In this chapter, we will focus on the synthesis, characterization, and diagnostic applications of nanoflares as they relate to cancer markers.
Subject(s)
Immobilized Nucleic Acids , Metal Nanoparticles , Nanoconjugates , Nanomedicine/methods , Neoplasms/diagnosis , Animals , Fluorescent Dyes , Humans , Immobilized Nucleic Acids/chemical synthesis , Immobilized Nucleic Acids/chemistry , Metal Nanoparticles/chemistry , Nanoconjugates/chemistryABSTRACT
This review focuses on surface-grafted DNA, and its use as a molecular building block that exploits its unique properties as a directional (poly)anion that exhibits molecular recognition. The selected examples highlight innovative applications of DNA at surfaces and interfaces ranging from molecular diagnostics and sequencing to biosensing.
